M1 macrophage-derived exosomes impair beta cell insulin secretion via miR-212-5p by targeting SIRT2 and inhibiting Akt/GSK-3β/β-catenin pathway in mice
M1 macrophage-derived exosomes impair beta cell insulin secretion via miR-212-5p by targeting SIRT2 and inhibiting Akt/GSK-3β/β-catenin pathway in mice
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DOI：
10.1007/s00125-021-05489-1
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发表时间：
2021-06-11
期刊：
影响因子：
8.2
通讯作者：
Han, Xiao
中科院分区：
文献类型：
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作者：
Qian, Bin;Yang, Yang;Han, Xiao
目的/假设胰岛中巨噬细胞水平升高，并且所产生的炎症反应是肥胖和2型糖尿病期间β细胞衰竭的主要原因。我们和其他人先前的研究已经报道，巨噬细胞释放的外泌体在介导细胞间通讯中起重要作用，并且代表了一类参与与2型糖尿病相关的炎症过程的炎症因子。然而，到目前为止，还没有报道证明巨噬细胞来源的外泌体对β细胞的作用，并且关于它们在β细胞损伤中的潜在机制知之甚少。因此，我们的目的是研究巨噬细胞衍生的外泌体对胰岛β细胞损伤的影响，在体外和体内invivo.Methods胰岛居民巨噬细胞的表型配置文件进行了分析，在C57 BL/6 J小鼠喂食高脂饮食（HFD）。从培养的骨髓来源的巨噬细胞（BMDM）的培养基和从HFD喂养的小鼠的分离的胰岛驻留巨噬细胞（HFD-Exos）收集外泌体。评估了由炎性M1表型BMDM（M1-Exos）和HFD-Exos分泌的外泌体对β细胞功能的作用。进行miRNA微阵列和定量实时PCR（qPCR）以测试β细胞中M1-Exos衍生的miR-212- 5 p的水平。然后，miR-212- 5 p在M1-Exos或β细胞中过表达或抑制，以确定其分子和功能impacts.Results M1极化的巨噬细胞在肥胖小鼠的胰岛中富集。HFD喂养小鼠的M1巨噬细胞和胰岛驻留巨噬细胞以外泌体依赖性方式损害β细胞胰岛素分泌。miR-212- 5 p在M1-Exos和HFD-Exos中显著上调。增加miR-212- 5 p的表达损害β细胞胰岛素分泌。阻断miR-2 12- 5 p引起损伤期间M1-Exos介导的β细胞胰岛素分泌的显著改善。从机制上讲，M1-Exos介导了miR-212- 5 p的细胞间转移，靶向沉默调节蛋白2基因，并调节受体β细胞中的Alct/GSK-3 β/β-连环蛋白途径以限制胰岛素分泌。结论/解释一种新的外泌体调节机制被描绘为巨噬细胞-β细胞串扰，驱动β细胞功能障碍，并应探索其治疗效用。
Aims/hypothesis Macrophage levels are elevated in pancreatic islets, and the resulting inflammatory response is a major contributor to beta cell failure during obesity and type 2 diabetes mellitus. Previous studies by us and others have reported that exosomes released by macrophages play important roles in mediating cell-to-cell communication, and represent a class of inflammatory factors involved in the inflammatory process associated with type 2 diabetes mellitus. However, to date, no reports have demonstrated the effect of macrophage-derived exosomes on beta cells, and little is known regarding their underlying mechanisms in beta cell injury. Thus, we aimed to study the impact of macrophage-derived exosomes on islet beta cell injury in vitro and in vivo.Methods The phenotypic profiles of islet-resident macrophages were analysed in C57BL/6J mice fed a high-fat diet (HFD). Exosomes were collected from the medium of cultured bone marrow-derived macrophages (BMDMs) and from isolated islet-resident macrophages of HFD-fed mice (HFD-Exos). The role of exosomes secreted by inflammatory M1 phenotype BMDMs (M1-Exos) and HFD-Exos on beta cell function was assessed. An miRNA microarray and quantitative real-time PCR (qPCR) were conducted to test the level of M1-Exos-derived miR-212-5p in beta cells. Then, MiR-212-5p was overexpressed or inhibited in M1-Exos or beta cells to determine its molecular and functional impact.Results M1-polarised macrophages were enriched in the islets of obese mice. M1 macrophages and islet-resident macrophages of HFD-fed mice impaired beta cell insulin secretion in an exosome-dependent manner. miR-212-5p was notably upregulated in M1-Exos and HFD-Exos. Enhancing the expression of miR-212-5p impaired beta cell insulin secretion. Blocking miR-2 12-5p elicited a significant improvement in M1-Exos-mediated beta cell insulin secretion during injury. Mechanistically, M1-Exos mediated an intercellular transfer of the miR-212-5p, targeting the sirtuin 2 gene and regulating the Alct/GSK-3 beta/beta-catenin pathway in recipient beta cells to restrict insulin secretion.Conclusions/interpretation A novel exosome-modulated mechanism was delineated for macrophage-beta cell crosstalk that drove beta cell dysfunction and should be explored for its therapeutic utility.